Articles
EARLY INVESTIGATION ON THE EFFECTS OF PVS2 DURATION FOR CRYOPRESERVATION OF SPATHOGLOTTIS PLICATA ORCHID PROTOCORMS
Article number
1039_21
Pages
167 – 172
Language
English
Abstract
Cryopreservation is an alternative and an important tool in promoting ex situ conservation of orchid germplasm.
This study was conducted to investigate the potential of cryopreserving the encapsulated and non-encapsulated protocorms of Spathoglottis plicata using plant vitrification solution 2 (PVS2). Protocorms (1-2 mm) were precultured with half strength modified Murashige and Skoog (MS) liquid medium supplemented with 0.3 M sucrose for 24 h followed by encapsulation with 2% sodium alginate and 0.3 M sucrose.
Both encapsulated and non-encapsulated non-cryopreserved protocorms were treated with a loading solution containing 2M glycerol with 0.4 M sucrose for 20 min and subsequently dehydrated with PVS2 to different exposure times (5, 10, 15, 20, 25 and 30 min) prior to liquid nitrogen (LN) storage determination.
The survival rate of the encapsulated protocorms in all PVS2 dehydration treatment was 36 to 52% with 6 to 36% developed into normal plantlets.
However, non-encapsulated protocorms only gave 3-7% survival rate at 5-15 min and 11% developed as normal plantlet at 10 min dehydration time.
Non-encapsulated protocorm did not survive after 20 min dehydration in PVS2 solution but encapsulated protocorm did survive.
Encapsulations using alginate solution enhanced protection to S. plicata protocorms and reduced the osmotic stress to PVS2 solution.
Hence, further investigation and optimization on other important parameters in encapsulated-vitirification technique should be carried out as it is necessary for Spathoglottis plicata cryopreservation.
This study was conducted to investigate the potential of cryopreserving the encapsulated and non-encapsulated protocorms of Spathoglottis plicata using plant vitrification solution 2 (PVS2). Protocorms (1-2 mm) were precultured with half strength modified Murashige and Skoog (MS) liquid medium supplemented with 0.3 M sucrose for 24 h followed by encapsulation with 2% sodium alginate and 0.3 M sucrose.
Both encapsulated and non-encapsulated non-cryopreserved protocorms were treated with a loading solution containing 2M glycerol with 0.4 M sucrose for 20 min and subsequently dehydrated with PVS2 to different exposure times (5, 10, 15, 20, 25 and 30 min) prior to liquid nitrogen (LN) storage determination.
The survival rate of the encapsulated protocorms in all PVS2 dehydration treatment was 36 to 52% with 6 to 36% developed into normal plantlets.
However, non-encapsulated protocorms only gave 3-7% survival rate at 5-15 min and 11% developed as normal plantlet at 10 min dehydration time.
Non-encapsulated protocorm did not survive after 20 min dehydration in PVS2 solution but encapsulated protocorm did survive.
Encapsulations using alginate solution enhanced protection to S. plicata protocorms and reduced the osmotic stress to PVS2 solution.
Hence, further investigation and optimization on other important parameters in encapsulated-vitirification technique should be carried out as it is necessary for Spathoglottis plicata cryopreservation.
Authors
F.C. Ginibun, R.Y. Othman, S. Bhassu , N. Khalid
Keywords
cryopreservation, Spathoglottis plicata, protocorm, PVS2, encapsulation-vitrification
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